CN102108090B - Dihydroiso-coumarin glucoside compounds and preparation method and application thereof - Google Patents
Dihydroiso-coumarin glucoside compounds and preparation method and application thereof Download PDFInfo
- Publication number
- CN102108090B CN102108090B CN 200910200557 CN200910200557A CN102108090B CN 102108090 B CN102108090 B CN 102108090B CN 200910200557 CN200910200557 CN 200910200557 CN 200910200557 A CN200910200557 A CN 200910200557A CN 102108090 B CN102108090 B CN 102108090B
- Authority
- CN
- China
- Prior art keywords
- water
- ethanol
- aqueous
- acetone
- column chromatography
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses new dihydroiso-coumarin glucoside compounds shown in a formula I, which are extracted and separated from Periplaneta americanna medicinal materials. Through mouse auricle swelling tests, rat granulation tissue hyperblastosis tests and the like, results show that the compounds have effects of resisting inflammation, promoting wounds to heal and the like; and the compounds can be used for preparing medicines for treating inflammation, ulcers, wounds, incised wounds, fistula, burn, scalding and the like, particularly medicines for preventing and treating diseases related to inflammation resistance, promotion of granulation tissue hyperblastosis, wound healing and the like, and have high clinical application value. The invention provides a preparation method for the compounds shown in the formula I.
Description
Technical field
The present invention relates to pharmaceutical chemistry, be specifically related to a kind of Dihydroiso-coumarin glucoside new compound that from periplaneta americana (Periplaneta americana) medicinal material, extracts separation and preparation method thereof and application.
Background technology
Periplaneta americana Periplaneta americana is Insecta Pterygota Blattodea Blattidae Periplaneta insect, is commonly called as " cockroach ", and it is used as medicine and begins to be stated from Shennong's Herbal, and it is classified as middle product, meaning " flavor: salty, cold; Control: the hard fever and chills of Blood stasis, brokenly gather, laryngopharynx closes, interior cold s.m.p "." cockroach " sees supplementary Amplifications of the Compendium of Materia Medica for its popular name head, is also referred to as stone ginger, sliding worm, and there is its different title method the each place, as steals oily mother-in-law, kitchen range ant etc.There have been 3.2 hundred million years in Blattidae Blattidae on earth as a huge insect family, belongs to the biology that the same epoch occur with dinosaur.It is the strongest, the most ancient, one of the most successful insect assembly of procreation so far of vitality in the world.Global blattaria is near 5000 kinds according to statistics at present, and wherein, China is recited as 253 kinds, mainly is distributed in the indoor or open-air of tropical and subtropical region.Periplaneta americana is one of a few Domestic Cockroaches kind.It mainly inhabits kitchen, dining room, warehouse etc. and locates to damage food as indoor pest, clothes, and books etc. take food and contaminated food simultaneously, propagate dysentery, typhoid fever, roundworm, the various diseases such as pinworm.(Chinese media biology and control magazine, 1999,10 (3): 168.) chamber occupies enteron aisle that blattaria may carry and respiratory tract malignant bacteria up to 34 kinds, 12 kinds in mould, and the multiple virus that comprises bone marrow poliomyelitis virus etc. for Hu Xiuyuan report.Yet in the face of various agent for killing, this insect colony is still very active, shows that its body planted agent has some special material and distinctive mechanism of action, and this has caused more domestic and international scholars' concern.From numerous research report documents to periplaneta americana, present report mainly concentrates on the distribution of various blattarias and kills situation report; Various agent for killing to its efficiency ratio of killing with kill research on mechanism; The research of its genetic expression and sensitization aspect.On the other hand, begin it is studied as medicine, to its chemical ingredients, comprise the extraction of various enzymes, peptide and protein, separate the research of evaluation and physiological function thereof and mechanism of action aspect etc.By existing product being reached relevant pharmacology activity research situation analysis, the biological activity of effective component mainly has anti-inflammatory, detumescence, promotion cell proliferation and new granulation tissue's growth in the periplaneta americana body, accelerate the disease damage tissue repair, accelerate the effects such as necrotic tissue comes off, various traumatic surfaces and body ulcer are had the active (product: Kangfuxin Liquid) of good tissue repair; Have boosting, improve sanguimotor activity (product: Xinmailong).Activity (product: liver is grand) with anti-bacteria and anti-virus; Has antitumor, enhancing immunologic function activity (product: eliminating mass benefit liver sheet) etc.But the said products is complicated because of its ingredient, and real active substance is still not clear.Chemical ingredients in the report periplaneta americana is mainly pheromone class and amino acid and various insect neuropeptide class at present, also have in addition blattaria acid, periplanone, cockroach oil, anaphylactogen composition, enzyme, CyA, B, C, chitin, nicotinic acid and trace element etc., but, have the report of clear and definite drug activity chemical ingredients less.Present disclosed patent mainly is extract, composition, does not illustrate its active substance basis from molecular level, can not be with clear and definite composition as quality control index.
The inventor has carried out deep chemical constitution study to American-cockroach-extract, and by active tracking screening, successfully be separated to the new compound monomer component that has significant anti-inflammatory, promotes granulation tissue hyperplasia, the effect of wound healing isoreactivity, and proved conclusively its chemical structure.
Summary of the invention
Technical problem to be solved by this invention is to research and develop the natural compounds composition with aspect pharmacologically actives such as anti-inflammatory, wound healings from periplaneta americana, for the preparation of having anti-inflammatory, promoting the pharmaceutical composition that granulation tissue hyperplasia, wound healing etc. act on.
The invention provides a kind of Dihydroiso-coumarin glucoside new compound, this compound has the chemical structural formula shown in the formula I:
Chemical name: (3R)-3-methyl-7-methylol-8-hydroxyl-3,4-Dihydroiso-coumarin-6-O-β-D-glucopyranoside
English name: pericanaside
Code name: KFX-4 (code name of inventor's appointment)
Have no by literature search report, be a kind of new compound, called after pericanaside.
Compound I is micro-yellow powder, soluble in water, ethanol, methyl alcohol etc.;
The high resolution electrospray ionization mass spectrum is measured molecular formula C
17H
22O
10, m/z 385.1139[M-H];
Calculated value 385.1135; Degree of unsaturation=7.
The UV spectrum absorption band is at λ max:221,264.7,306.2nm.
There is hydroxyl (v3416cm-1, strong broad peak) in the infrared spectra prompting structure, ester group (v1659cm-1) and aromatic ring v1625,1588cm-1).
Compound I
1The HNMR collection of illustrative plates discloses: δ 2.98 (1H, dd, J=16.4Hz; 3.6Hz), 2.89 (1H, dd, J=16.4Hz; 11.2Hz) and δ 4.75 (1H, m) be an ABX spin system; H-HCOSY spectrum δ 1.42 (3H, d, J=6.0Hz) is relevant with δ 4.75 (1H, m).δ 4.50 (2H, ABq, J=11.2Hz) is that (signal CH2-OH), two hydrogen of methylol cause its chemical shift non-equivalence because chemical environment of living in is different to a methylol, show as the AB spin system.δ 6.64 (1H, s) is isolated fragrant hydrogen signal.There are the phenolic hydroxyl group with the carbonyl chelating, i.e. a C-8 hydroxyl in δ 11.43 (1H, br) the signal prompt structure.
13CNMR: δ 101.76,74.20,77.07,70.35, have a glucopyranose units in the 78.16 and 61.40 signal prompt structures, consider 1HNMR δ 4.90 (1H, d, J=7.20Hz), and this glucose unit should be β-D-glucopyranoside.13CNMR shows 17 peaks, and wherein δ 170.50 is the carbonyl signal, 21.10,3 CH2 of a methyl δ, 7 CH, 5 quaternary carbon signals from the phenyl ring contribution.HMBC discloses: H-9 and C-3, C-4 exist distant relation H-5 and C-4, C-6, and C-8a is relevant; H-4 and C-3, C-8a is relevant; H-9 is relevant with C-6 with H-1 '.The stereochemistry of C-3, two on the C-4 show as the AB spin system with hydrocarbonize displacement study non-equivalence with the carbon coupling, and these two hydrogen form respectively the AB part of ABX spin system with the H-3 coupling.By above Spectral Characteristic inferred from input data, this compound should be 3-methyl-8-hydroxyl-3,4-Dihydroiso-coumarin glucoside compounds.
Table 1
1H and
13CNMR Compound I (pericanaside)
| Positio n | δH | COSY | δC | DEPT | HMBC |
| 1 | 170.50 | |
|||
| 2 | |||||
| 3 | 4.75(1H，m) | H-9，H-4 | 76.52 | |
|
| 4 | 2.98(1H，dd，J＝16.4；3.6) 2.89(1H，dd，J＝16.4；11. 2) | H-3 | 34.76 | CH2 | |
| 4a | 142.09 | |
|||
| 5 | 6.64(1H，s) | 116.97 | |
||
| 6 | 162.6 | C | 34.76，162.24 | ||
| 7 | 106.39 | |
|||
| 8 | 161.60 | C | |||
| 8a | 103.37 | |
|||
| 9 | 1.42(3H，d，J＝6.0) | 21.10 | CH3 | 76.5，34.76 | |
| 10 | 4.50(2H，ABq，J＝11.2 | 52.08 | CH2 | 116.97， 116.60 | |
| 1’ | 4.90(1H，d，J＝7.20) | H-2’ | 101.76 | CH | 162.24 |
| 2’ | 3.30 | 74.20 | CH | ||
| 3’ | 3.27(1H，t，J＝8.4) | 77.07 | CH | ||
| 4’ | 3.19(1H，t，J＝8.8) | 70.35 | CH | ||
| 5’ | 3.35(m) | 78.16 | CH | ||
| 6’ | 3.70(1H，dd，J＝11.6；3.2) 3.49(1H，dt，J＝11.6；5.6 | H-6’ H-5’ | 61.40 | CH2 | 78.16 |
| 2’-OH | 5.44(1H，br.s) | ||||
| 3’-OH | 5.15(1H，br.s) | ||||
| 4’-OH | 5.10(1H，br.s) | ||||
| 6’-OH | 4.62(1H，t，J＝5.2) | ||||
| 8-OH | 11.43(1H，br) |
Assignments were made by1D and 2DNMR 1H-1H-COSY，HSQC，HMBC，andNOE experiments；δin ppm，J in Hz.
NMR Experiments：with a Bruker DPR-400 spectrometer，at 400(1H)and 100MHz(13C)；chemical shifts δin ppm rel.to DMSO-d6(2.50，1H；40.35，13C)
Another object of the present invention has provided the preparation method of above-mentioned a kind of Dihydroiso-coumarin glucoside compounds, and the method comprises the following steps:
(1) extract:
Dry or bright adult is raw material with Blattidae insect periplaneta americana Periplaneta americana, with solvent extraction 1-3 time;
(2) extraction: the concentrated solution that said extracted liquid concentrating under reduced pressure reclaims behind the solvent adds water extraction 1-3 time, and the consumption of water is the 2-4 doubly (w/w) of concentrated solution, fully stirs standing demix, water intaking layer after adding water;
(3) absorption: water layer is through macroporous adsorbent resin or charcoal absorption, adsorptive elutriant wash-out;
(4) separate: the elutriant silica gel column chromatography, one or more modes of LH-20 column chromatography or reversed phase column chromatography method or preparation HPLC method are separated, water, ethanol, methyl alcohol, propyl carbinol, acetone, acetonitrile, ethyl acetate or sherwood oil or their at least two kinds of mixtures are made the eluent wash-out, adopt thin layer chromatography or UV to detect, merge the elutriant that contains target compound, behind 50-70 ℃ of vacuum lyophilization or the vacuum heat drying, get target compound (3R)-3-methyl-7-methylol-8-hydroxyl-3,4-Dihydroiso-coumarin-6-O-β-D-glucopyranoside.
The described step of the inventive method (1) is when extracting raw material, in normal temperature or heating, under normal pressure or the decompression state, take the aqueous solution of ethanol, methyl alcohol, acetone or the propyl alcohol of 20%-100% as solvent extraction; Each consumption that extracts solvent is 4-10 times of w/w of raw material; Extracting method is decoction, reflux, supersound extraction, cold soaking, diacolation or microwave extraction, and preferred 50-95% ethanol heating is extracted 1-3 time; Each 3-12 of extraction time hour.
The described step of the inventive method (3) macroporous adsorbent resin is the polystyrene type polymeric adsorbent, comprise D101, MN200, HPD100, HPD400, HPD450, AB-8, D4020, S-8, HZ806 or HP-20, preferred D101, MN200 or HPD100 type macroporous adsorbent resin; Eluting solvent is water, aqueous ethanol, aqueous methanol, aqueous acetone or moisture propyl alcohol, and the water content of water-containing solvent is at 5%-95%v/v; Elution process is isocratic elution or gradient elution.
When described step (3) adopts Flavonoids by Macroporous Adsorption Resin, first with the washing of 1-8 times of column volume; Use or without the aqueous cleaning of ethanol, methyl alcohol, acetone or the propyl alcohol of 5%-20%, further remove impurity, consumption is 1-3 times of column volume; Use at last the aqueous solution wash-out of ethanol, methyl alcohol, acetone or the propyl alcohol of 20%-95%, consumption is 2-8 times of column volume, collects elutriant, reclaims solvent, namely get and contain (3R)-3-methyl-7-methylol-8-hydroxyl-3, the part of 4-Dihydroiso-coumarin-6-O-β-D-glucopyranoside.It is powdered carbon or granular active carbon that described step (3) adopts the gac of active carbon adsorption; Eluting solvent is water, aqueous ethanol, aqueous methanol, aqueous acetone or moisture propyl alcohol, and the water content of water-containing solvent is at 5%-95%; Elution process is isocratic elution or gradient elution.
During described step (3) charcoal absorption, first with 1-8 times of column volume washing; Use or without the aqueous cleaning of ethanol, methyl alcohol, acetone or the propyl alcohol of 5%-20%, further remove impurity, consumption is 1-3 times of column volume; Use at last the aqueous solution wash-out of ethanol, methyl alcohol, acetone or the propyl alcohol of 20%-95%, consumption is 2-8 times of column volume, collects elutriant, reclaims solvent, namely get and contain (3R)-3-methyl-7-methylol-8-hydroxyl-3, the part of 4-Dihydroiso-coumarin-6-O-β-D-glucopyranoside.
The used silica gel column chromatography of the described step of the inventive method (4) column chromatography is normal pressure or pressurized column chromatography, used filler is 100-400 order silica gel, be eluent with ethyl acetate, propyl carbinol, water, propyl alcohol, ethanol, methyl alcohol, acetone, sherwood oil or normal hexane or their at least two kinds of mixtures, elution process is isocratic elution or gradient elution;
When described step (4) adopts sephadex LH-20 column chromatography, adopt normal pressure or pressurized column chromatography, used filler is sephadex LH-20 dextrane gel, adopting the aqueous solution of ethanol, methyl alcohol, acetone or propyl alcohol is eluent, the moisture 10%-90%v/v of eluent, elution process are isocratic elution or gradient elution;
When described step (4) adopts the reversed phase column chromatography method, adopt normal pressure or pressurized column chromatography or preparation HPLC method, used filler is Octadecylsilane bonded phase or eight alkyl bonded phases, in employing aqueous ethanol, aqueous methanol, aqueous acetone or the moisture propyl alcohol one or more are eluent, eluent moisture ratio 10%-90%v/v, elution process are isocratic elution or gradient elution.
Another purpose of the present invention has provided the application of above-mentioned a kind of Dihydroiso-coumarin glucoside compounds in the preparation medicine.
Anti-inflammatory activity test (p-Xylol causes the impact of mice ear) result and the model control group of the compounds of this invention I compare, and KFX-4 is low, high dose group obviously reduces (P＜0.01), 62.5mgkg than the ear swelling degree of positive controls mouse
-1The antiphlogistic effects of KFX-4 and 25mgkg
-1Hydrocortisone is suitable; Preliminary experiment results shows, but the mice auricle swelling due to the KFX-4 establishment dimethylbenzene shows that KFX-4 has obvious anti-inflammatory action.
The promotion granulation tissue hyperplasia test of the compounds of this invention, the result shows and blank the group relatively that granuloma weight in wet base and the dry weight of KFX-4 group rat obviously increase (P＜0.01).Pathological examination results is respectively organized sample standard deviation and is seen new granulation tissue and foreign matter granulation tissue, also see the local necrosis tissue, a large amount of fusiformis inoblasts in the granulation tissue, local visible capillary vessel and foreign matter scavenger cell, and the inflammatory cell infiltrations such as lymphocyte and neutrophilic granulocyte are arranged.Test-results shows that KFX-4 can significantly promote granulation tissue hyperplasia.
Above-mentioned test-results shows that the compounds of this invention can be used for preparation prevention and the medicines such as treatment inflammation, ulcer, wound, metal-inflicted wound, fistula, burn, scald, especially prepares the medicine of the relevant disease such as prevention and treatment and anti-inflammatory, promotion granulation tissue hyperplasia, wound healing.The pharmaceutical composition that described medicine is made as activeconstituents and pharmaceutical excipient by Dihydroiso-coumarin glucoside compounds of the present invention.Comprise mixture, oral liquid, gelifying agent, ointment, capsule, tablet, granule and injection etc.
Description of drawings
Fig. 1. the uv-absorbing figure of compound 1
Fig. 2. the infrared spectrogram of compound 1
Fig. 3. the high resolution electrospray ionization mass spectrum figure of compound 1
Fig. 4. the 1H-NMR figure of compound 1
Fig. 5. the 1H-NMR partial enlarged drawing (a) of compound 1
Fig. 6. the 1H-NMR partial enlarged drawing (b) of compound 1
Fig. 7. the 13C-NMR figure of compound 1
Fig. 8. the DEPT135 figure of compound 1
Fig. 9. the DEPT90 figure of compound 1
Figure 10. the DEPT45 figure of compound 1
Figure 11. the 1H-1H COSY figure of compound 1
Figure 12. the HMBC figure of compound 1
Figure 13. the HSQC figure of compound 1
Figure 14. the NOESY figure of compound 1
Figure 15. the CD of compound 1 (circular dichroism spectrum) figure
Embodiment
Below in conjunction with specific embodiment, the invention will be further described, but content of the present invention is not limited only to cited embodiment.
The dry adult 20kg of periplaneta americana (Periplaneta americana) gets 3 times with 60 ℃ of warm lixiviates of 95% ethanol, and each consumption is respectively 160kg, 120kg, 120kg, each 12 hours of extraction time.(60 ℃ of extracting solution concentrating under reduced pressure,-0.08MPa) reclaim solvent to distinguishing the flavor of without alcohol, get concentrated solution 10kg, add the dissolving of 20kg-40kg deionized water and stirring, leave standstill and make layering, water intaking layer, (60 ℃ of concentrating under reduced pressure,-0.08MPa) to thick paste (relative density 1.20-1.30), namely get American-cockroach-extract (containing new compound I) 1.35kg.
The dry adult 10kg of periplaneta americana (Periplaneta americana) extracts 3 times with 50% alcohol heating reflux, and each consumption is respectively 100kg, 80kg, 60kg, each 3 hours of extraction time.(60 ℃ of extracting solution concentrating under reduced pressure,-0.08MPa) reclaim solvent to distinguishing the flavor of without alcohol, get concentrated solution 5kg, add the dissolving of 20kg deionized water and stirring, leave standstill and make layering, water intaking layer, (60 ℃ of concentrating under reduced pressure,-0.08MPa) to thick paste (relative density 1.20-1.30), namely get American-cockroach-extract (containing new compound I) 740g.
Get the American-cockroach-extract 300g that embodiment 1 (or embodiment 2) extracts, add water 900ml dissolving, upper D101 macroporous adsorbent resin (consumption 1kg, also can be with other model such as MN200, HPD100, HPD400, HPD450, AB-8, D4020, S-8, HZ806, the resins such as HP-20), loading finishes, adsorptive is used first the washing of 5 times of column volumes, aqueous ethanolic solution with 2 times of column volumes 10% cleans removal impurity again, use again the aqueous ethanolic solution wash-out of 5 times of column volumes 30%, collect 30% alcohol elution, reclaim solvent, namely get the part that contains KFX-4.Again through silica gel column chromatography, blade diameter length ratio 1: 50, silica gel (300-400 order) consumption 100g, eluent: propyl carbinol: ethyl acetate: water=3; 12: 1; Every 20ml collects 1 stream part, and the TLC method detects each stream part (developping agent: propyl carbinol: ethyl acetate: water=3; 6: 1; Developer: 10%F
eCl
3Test solution), collect KFX-4 place stream part, after merging KFX-4 place stream part, 60 ℃ are evaporated to 3-5ml, upper LH-20 column chromatography, methyl alcohol: water=1: 1 wash-out, collect and flow part, the TLC method detects each stream part, stream part of the single spot of merging KFX-4, reclaim solvent, freeze-drying namely gets new compound I (61mg).
Get the American-cockroach-extract 200g that embodiment 1 (or embodiment 2) extracts, add water 600ml dissolving, upper D101 absorption with macroporous adsorbent resin (consumption 600g), loading finishes, and adsorptive is used first the washing of 5 times of column volumes, and the aqueous ethanolic solution with 2 times of column volumes 10% cleans removal impurity again, use again the aqueous ethanolic solution wash-out of 5 times of column volumes 40%, collect 40% alcohol elution, reclaim solvent, namely get the part that contains KFX-4.Go up again the LH-20 column chromatography, methyl alcohol: water=1: 1 wash-out, to collect and flow part, the TLC method detects respectively stream part, and merging KFX-4 place stream part is reclaimed solvent and is the KFX-4 crude product; Crude product is again through ODS column chromatography (preparation HPLC), moving phase: methyl alcohol: water=1: 3, UV265nm detects, and collects KFX-4 peak stream part, reclaim solvent after, freeze-drying namely gets new compound I (40mg).
Get the American-cockroach-extract 200g that embodiment 1 (or embodiment 2) extracts, add water 600ml dissolving, upper MN200 macroporous adsorbent resin (consumption 600g), loading finishes, first with 6 times of column volume washings, aqueous ethanolic solution with 2 times of column volumes 20% cleans removal impurity again, uses the aqueous ethanolic solution wash-out of 5 times of column volumes 40% again, collects 40% alcohol elution, reclaim solvent, namely get the part that contains KFX-4, upper LH-20 column chromatography, methyl alcohol: water=1: 1 wash-out, collect stream part, the TLC method detects respectively stream part, merges to contain KFX-4 stream part, reclaims solvent and is the KFX-4 crude product; Crude product is again through ODS column chromatography (preparation HPLC), moving phase: methyl alcohol: water=1: 3, UV265nm detects, and collects KFX-4 peak stream part, reclaim solvent after, freeze-drying namely gets new compound I (37mg).
Get the American-cockroach-extract 200g that embodiment 2 (or embodiment 1) extracts, add water 500ml dissolving, add gac 100g, stirring makes abundant absorption, and the dress post is first with 5 times of column volume deionization washings, use again 5 times of column volume 95% ethanol elutions, collect 95% ethanol eluate, Recycled ethanol namely gets the part that contains KFX-4.Upper LH-20 column chromatography, methyl alcohol: water=1: 2 wash-out, collect stream part, the TLC method detects each stream part, merges KFX-4 place stream part, concentrating under reduced pressure (60 ℃ ,-0.08MPa) after again through silica gel column chromatography, eluent: propyl carbinol: ethyl acetate: water=3; 12: 1; Collect and flow part, the TLC method detects each stream part, and stream part of the single spot of merging KFX-4 is reclaimed solvent, is drying to obtain new compound I (35mg).
Get the American-cockroach-extract 100g that embodiment 1 (or embodiment 2) extracts, add water 300ml dissolving, upper HPD100 absorption with macroporous adsorbent resin (consumption 400g), loading finishes, remove impurity with 8 times of column volume washings first, use again 6 times of column volumes, 50% aqueous ethanolic solution wash-out, collect 50% alcohol elution, reclaim solvent, namely get the part that contains KFX-4, upper LH-20 column chromatography, ethanol: water=1: 1 wash-out, collect successively stream part, every part of 10ml, the TLC method detects respectively stream part, merges to contain KFX-4 stream part, reclaims solvent and is the KFX-4 crude product; Crude product is again through ODS column chromatography (preparation HPLC), and moving phase: methyl alcohol: water=1: 3, UV265nm detects, and collects KFX-4 peak stream part, behind the recovery solvent, is drying to obtain new compound I (20mg).
KFX-4 (embodiment 5 preparations) p-Xylol causes the impact of mice ear
Get 50 of the male ICR mouses of 21～23g, divide at random 5 groups by body weight: Normal group, model control group, KFX-4 low dose group (KFX-4 25mgkg
-1), KFX-4 high dose group (KFX-4 62.5mgkg
-1), positive controls (hydrocortisone 25mgkg
-1), every group 10, each organizes 30min behind the corresponding medicine of abdominal injection and the solvent, except Normal group, get 30 μ l dimethylbenzene and evenly be applied in the left ear of mouse (square two sides each 15 μ L), cause scorching 45min after, take off mouse left and right sides ear with punch tool, and accurately weigh with analytical balance, calculate the mice ear degree: (left ear-auris dextra)/auris dextra.
Table KFX-4 p-Xylol causes the impact (X ± s, n=10) of mice ear
Annotate: compare with Normal group,
△P＜0.05,
△ △P＜0.01; With the model control group comparative group,
*P＜0.05,
*P＜0.01.
The ear swelling degree of, high dose group low with model control group comparison KFX-4 and positive controls mouse obviously reduces (P＜0.01), 62.5mgkg
-1The antiphlogistic effects of KFX-4 and 25mgkg
-1Hydrocortisone is suitable; Preliminary experiment results shows, but the mice auricle swelling due to the KFX-4 establishment dimethylbenzene shows that KFX-4 has obvious anti-inflammatory action.
KFX-4 (embodiment 3 preparations) promotes the granulation tissue hyperplasia test
Get body weight and be 20 of the male SD rats of 270～300g, divide at random 2 groups by body weight: blank group, KFX-4 group, each organize rat with 3% vetanarcol (45mg/kg) intraperitoneal anesthesia after, rat back is shaved hair with shave, and uses iodophor disinfection, under aseptic condition, the subcutaneous placement plastic hoop, ring is sewed up a pin with the skin contact place, and collar extension is exposed to outside the skin, is open.Performed the operation rear 1 day, each is organized rat and splashes into medicine or the solvent 0.5ml of corresponding concentration, every twice-daily at the plastic hoop opening part, each 0.25ml, successive administration 10 days behind the last administration 24h, is put to death each treated animal, get granulation tissue, blot tissue juice with filter paper, and weigh with analytical balance, get and measure dry weight after half granuloma places 65 ℃ of baking oven baking 24h, second half granuloma is fixedly carried out pathologic finding with formaldehyde, and whether observe is granulation tissue.
Table KFX-4 is on the granulomatous impact of rat (X+s, n=10)
Annotate: compare with the blank group,
*P＜0.05,
*P＜0.01.
By table 1 result as seen, compare with the blank group, granuloma weight in wet base and the dry weight of KFX-4 group rat obviously increase (P＜0.01).Pathological examination results is respectively organized sample standard deviation and is seen new granulation tissue and foreign matter granulation tissue, also see the local necrosis tissue, a large amount of fusiformis inoblasts in the granulation tissue, local visible capillary vessel and foreign matter scavenger cell, and the inflammatory cell infiltrations such as lymphocyte and neutrophilic granulocyte are arranged.
Test-results shows that KFX-4 can significantly promote granulation tissue hyperplasia.
Claims (11)
1. Dihydroiso-coumarin glucoside compounds is characterized in that this compound has the chemical structural formula shown in the formula I:
2. the preparation method of Dihydroiso-coumarin glucoside compounds I as claimed in claim 1 is characterized in that the method comprises the following steps:
(1) extract:
Dry or bright adult is raw material with Blattidae insect periplaneta americana Periplaneta americana, in normal temperature or heating, under normal pressure or the decompression state, take the aqueous solution of ethanol, methyl alcohol, acetone or the propyl alcohol of 20%-100% as solvent extraction 1-3 time; Each consumption that extracts solvent is 4-10 times of w/w of raw material; Extracting method is that decoction, reflux, supersound extraction, cold soaking, temperature are soaked, diacolation or microwave extraction;
(2) extraction: the concentrated solution that said extracted liquid concentrating under reduced pressure reclaims behind the solvent adds water extraction 1-3 time, and the consumption of water is 2-4 times of w/w of concentrated solution, fully stirs standing demix, water intaking layer after adding water;
(3) absorption: water layer is through macroporous adsorbent resin or charcoal absorption, adsorptive elutriant wash-out; Described macroporous adsorbent resin is D101, MN200, HPD100, HPD400, HPD450, AB-8, D4020, S-8, HZ806 or HP-20 macroporous adsorbent resin; Eluting solvent is water, aqueous ethanol, aqueous methanol, aqueous acetone or moisture propyl alcohol, and the water content of water-containing solvent is at 5%-95%; Elution process is isocratic elution or gradient elution;
(4) separate: the elutriant silica gel column chromatography, one or more modes of LH-20 column chromatography or reversed phase column chromatography method or preparation HPLC method are separated, water, ethanol, methyl alcohol, propyl carbinol, acetone, acetonitrile, ethyl acetate or sherwood oil or their at least two kinds of mixtures are made the eluent wash-out, adopt thin layer chromatography or UV to detect, merge the elutriant that contains target compound, behind 50-70 ℃ of vacuum lyophilization or the vacuum heat drying, get target compound (3R)-3-methyl-7-methylol-8-hydroxyl-3,4-Dihydroiso-coumarin-6-O-β-D-glucopyranoside.
3. according to claim 2 method is characterized in that described step (1) when raw material is extracted, and in normal temperature or heating, under normal pressure or the decompression state, extracts 1-3 time take the aqueous solution of 50-95% ethanol as the solvent heating; Each 3-12 of extraction time hour.
4. according to claim 2 method is characterized in that described step (3) macroporous adsorbent resin is D101, MN200 or HPD100 type macroporous adsorbent resin.
5. according to claim 4 method is characterized in that described step (3) crosses macroporous adsorbent resin, first with the washing of 1-8 times of column volume; Use or without the aqueous cleaning of ethanol, methyl alcohol, acetone or the propyl alcohol of 5%-20%, further remove impurity, consumption is 1-3 times of column volume; Use at last the aqueous solution wash-out of ethanol, methyl alcohol, acetone or the propyl alcohol of 20%-95%, consumption is 2-8 times of column volume, collects elutriant, reclaims solvent, namely get and contain (3R)-3-methyl-7-methylol-8-hydroxyl-3, the part of 4-Dihydroiso-coumarin-6-O-β-D-glucopyranoside.
6. according to claim 2 method is characterized in that the gac of described step (3) active carbon adsorption is powdered carbon or granular active carbon; Eluting solvent is water, aqueous ethanol, aqueous methanol, aqueous acetone or moisture propyl alcohol, and the water content of water-containing solvent is at 5%-95%, and elution process is isocratic elution or gradient elution.
7. according to claim 6 method when it is characterized in that described step (3) charcoal absorption, washes first 1-8 times of column volume with water; Use or without the aqueous cleaning of ethanol, methyl alcohol, acetone or the propyl alcohol of 5%-20%, further remove impurity, consumption is 1-3 times of column volume v/v; Use at last the aqueous solution wash-out of ethanol, methyl alcohol, acetone or the propyl alcohol of 20%-95%, consumption is 2-8 times of column volume v/v, collect elutriant, reclaim solvent, namely get and contain (3R)-3-methyl-7-methylol-8-hydroxyl-3, the part of 4-Dihydroiso-coumarin-6-O-β-D-glucopyranoside.
8. according to claim 2 method, it is characterized in that the used silica gel column chromatography of described step (4) column chromatography is normal pressure or pressurized column chromatography, used filler is 100-400 order silica gel, be eluent with ethyl acetate, propyl carbinol, water, propyl alcohol, ethanol, methyl alcohol, acetone, sherwood oil or normal hexane or their at least two kinds of mixtures, elution process is isocratic elution or gradient elution; Or described step (4) LH-20 column chromatography, adopt normal pressure or pressurized column chromatography, used filler is sephadex LH-20 dextrane gel, adopting the aqueous solution of ethanol, methyl alcohol, acetone or propyl alcohol is eluent, the moisture 10%-90%v/v of eluent, elution process are isocratic elution or gradient elution; Described step (4) reversed phase column chromatography method, adopt normal pressure or pressurized column chromatography or preparation HPLC method, used filler is Octadecylsilane bonded phase or eight alkyl bonded phases, in employing aqueous ethanol, aqueous methanol, aqueous acetone or the moisture propyl alcohol one or more are eluent, eluent moisture ratio 10%-90%v/v, elution process are isocratic elution or gradient elution.
9. the as claimed in claim 1 application of Dihydroiso-coumarin glucoside compounds in the medicine of preparation prevention and treatment and anti-inflammatory, promotion granulation tissue hyperplasia or wound healing diseases related.
10. according to claim 9 application is characterized in that the described application that is applied as in the medicine of preparation prevention and treatment inflammation, ulcer, wound, metal-inflicted wound, fistula, burn or scald.
11. application according to claim 9, it is characterized in that described medicine is to contain the pharmaceutical composition that right requires 1 described Dihydroiso-coumarin glucoside compounds to make as activeconstituents and pharmaceutical excipient, comprises mixture, oral liquid, gelifying agent, ointment, capsule, tablet, granule and injection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200910200557 CN102108090B (en) | 2009-12-23 | 2009-12-23 | Dihydroiso-coumarin glucoside compounds and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200910200557 CN102108090B (en) | 2009-12-23 | 2009-12-23 | Dihydroiso-coumarin glucoside compounds and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102108090A CN102108090A (en) | 2011-06-29 |
| CN102108090B true CN102108090B (en) | 2013-04-03 |
Family
ID=44172382
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200910200557 Active CN102108090B (en) | 2009-12-23 | 2009-12-23 | Dihydroiso-coumarin glucoside compounds and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102108090B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102416023B (en) * | 2011-12-09 | 2013-03-20 | 浙江京新药业股份有限公司 | Periplaneta Americana L. gel externally applied medicinal composition and preparation method and use thereof in preparation of externally applied medicines for treating scalding |
| CN104622910A (en) * | 2013-11-08 | 2015-05-20 | 成都百草和济科技有限公司 | Low-temperature preparation method for American cockroach extract |
| CN107929299B (en) * | 2016-10-13 | 2020-10-16 | 内蒙古京新药业有限公司 | Antibacterial application of dihydroisocoumarin glucoside compound separated from periplaneta americana extract |
| CN107951882A (en) * | 2016-10-17 | 2018-04-24 | 内蒙古京新药业有限公司 | The pharmaceutical use of the isocoumarin class compound obtained is separated from American-cockroach-extract |
| CN108014137A (en) * | 2016-11-02 | 2018-05-11 | 浙江京新药业股份有限公司 | American-cockroach-extract lozenge |
| CN107929325B (en) * | 2017-11-29 | 2023-03-24 | 四川好医生攀西药业有限责任公司 | Suppository containing periplaneta americana and preparation method thereof |
| CN110672737B (en) * | 2019-09-29 | 2022-07-12 | 浙江京新药业股份有限公司 | Method for identifying periplaneta americana |
| CN114146097B (en) * | 2022-01-21 | 2023-06-23 | 大理大学 | Application of periplaneta americana extract and/or monomer in preparation of tumor microenvironment M2 polarization inhibition drugs |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1939345A (en) * | 2006-09-30 | 2007-04-04 | 耿福能 | American falx extract, medicinal composition containing it and its making method |
| CN1961893A (en) * | 2005-11-08 | 2007-05-16 | 肖小芹 | Method for preparing extract of adult bombay canary |
| CN101214262A (en) * | 2008-01-09 | 2008-07-09 | 大理学院 | American cockroaches effective fraction for anti-tumor prepared by macroporous adsorption resin and use |
-
2009
- 2009-12-23 CN CN 200910200557 patent/CN102108090B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1961893A (en) * | 2005-11-08 | 2007-05-16 | 肖小芹 | Method for preparing extract of adult bombay canary |
| CN1939345A (en) * | 2006-09-30 | 2007-04-04 | 耿福能 | American falx extract, medicinal composition containing it and its making method |
| CN101214262A (en) * | 2008-01-09 | 2008-07-09 | 大理学院 | American cockroaches effective fraction for anti-tumor prepared by macroporous adsorption resin and use |
Non-Patent Citations (2)
| Title |
|---|
| 美洲大蠊提取物抗炎、镇痛作用的实验研究;肖小芹等;《中国病原生物学杂志》;20070430(第02期);140-143 * |
| 肖小芹等.美洲大蠊提取物抗炎、镇痛作用的实验研究.《中国病原生物学杂志》.2007,(第02期), |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102108090A (en) | 2011-06-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102108090B (en) | Dihydroiso-coumarin glucoside compounds and preparation method and application thereof | |
| RU2349337C2 (en) | Pharmaceutical composition including steroid saponins, method of obtainment, and application | |
| Yao et al. | Anti-inflammatory diarylheptanoids and phenolics from the rhizomes of kencur (Kaempferia galanga L.) | |
| Han et al. | Protective effect of a polysaccharide from Rhizoma Atractylodis Macrocephalae on acute liver injury in mice | |
| Zhu et al. | Antipyretic and antitumor effects of a purified polysaccharide from aerial parts of Tetrastigma hemsleyanum | |
| Zhang et al. | Extremely low bioavailability of magnesium lithospermate B, an active component from Salvia miltiorrhiza, in rat | |
| KR101062670B1 (en) | Composition for the prevention or treatment of obesity-related diseases mediated by the activation of AMPK containing 2,5-bis-aryl-3,4-dimethyltetrahydrofuran lignan as an active ingredient | |
| Kong et al. | Optimization of extraction flavonoids from Exocarpium Citri Grandis and evaluation its hypoglycemic and hypolipidemic activities | |
| Niu et al. | Main iridoid glycosides and HPLC/DAD-Q-TOF-MS/MS profile of glycosides from the antioxidant extract of Eucommia ulmoides Oliver seeds | |
| Jia et al. | Hypoglycemic activity of Codonopsis pilosula (Franch.) Nannf. in vitro and in vivo and its chemical composition identification by UPLC-Triple-TOF-MS/MS | |
| Han et al. | Hepatoprotective glucosyloxybenzyl 2-hydroxy-2-isobutylsuccinates from Pleione yunnanensis | |
| Qiu et al. | Isolation, structural and bioactivities of polysaccharides from Anoectochilus roxburghii (Wall.) Lindl.: A review | |
| Zhong et al. | Qihuzha granule attenuated LPS-induced acute spleen injury in mice via Src/MAPK/Stat3 signal pathway | |
| Zhang et al. | A review of the traditional uses, phytochemistry, pharmacology and quality control of the ethnic medicinal plant Persicaria orientalis (L.) Spach in China | |
| CN103880913B (en) | A kind of compound and application thereof with hepatoprotective effect | |
| CN100591334C (en) | Usage of eleutheroside with anti-inflammatory action or aglycone thereof | |
| CN114409666A (en) | A homoisoflavonopolignan compound separated from fructus Hippophae, and its application in preparing medicine for treating non-alcoholic fatty liver disease | |
| Gou et al. | Traditional uses, phytochemistry, and pharmacology of Persicaria orientalis (L.) Spach-A review | |
| CN112939912A (en) | Preparation method and application of lactucin extracted from cichorium intybus | |
| CN113248483A (en) | Preparation method and application of lawn pennywort herb flavone glycoside monomer isovitexin | |
| CN111920799A (en) | Kulecuo effective component composition and preparation method and application thereof | |
| KR101402058B1 (en) | Composition comprising extract of fermented Curcuma longa by natural fermented soybean and curcuminoid derivatives isolated therefrom for treating or preventing liver disease | |
| CN103923156B (en) | There is saponin compound and the application thereof of hepatoprotective effect | |
| KR20020094725A (en) | Novel acid polysaccharide having anti-cancer immunoregulatory effect which is purified from red ginseng and composition for anti-cancer immunoreguation comprising same | |
| CN103864883B (en) | There is the triterpene saponin compound of liver protection function |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |